組織工程（tissue engineering）是21世紀十大熱門工作的榜首，藉由結合多種專業領域: 分子生物學、生物化學、材料科學、臨床醫學、生物醫學工程，組織工程的技術已凸顯其潛力以恢復組織或器官的缺損。 生物力學乃是組織再生中不可或缺的一環，目前細胞生物力學研究主要著重於研究細胞如何感應外在力學變化，進而轉換成細胞內訊號傳遞因子進行訊息傳導，此類藉由力學產生的訊號傳導又稱為“力學訊息傳遞 (Mechanotransduction)”，其可以調控細胞內基因使其進行細胞或組織重塑 (remodeling) 以影響其再生。

　　由於現代微機電系統 (MEMS) 及奈米科技的蓬勃發展，科學家已能達到微奈米 (微米: 10-6米; 奈米: 10-9米) 的操作水準進行細胞或次細胞級研究。本篇論文主要著重於結合生物微機電系統及奈米光電科技，建立多種操控及量測系統，以揭示細胞如何意識到機械刺激以及其細胞內訊號傳遞，進而量化因力學造成細胞基因調控的變化。

　　本研究利用雷射鑷系統並整合自製細胞刮取探針進行光學夾取及操作，量化量測細胞在不同粘附或移動時期的粘著力量變化。此外，藉由光罩製成技術 (microlithography)，我們將細胞外基質轉印成不同幾何形狀藉以控制細胞形狀或幾何限制細胞生長方向，然後施予不同方向的外力，以研究計畫性細胞凋亡 (Apoptosis) 和其挽救機制。為了即時量測細胞內分子活動，本論文亦成功架設全反射螢光顯微鏡(TIRFM)及螢光共振能量轉移系統 (FRET)，以量化探討施力方向對不同幾何外型的細胞其細胞粘著激脢 (FAK)、黏著相關分子 (Src)、以及細胞內骨骼 (Cytoskeleton)影響。

　　在探討外力對細胞基因調控層面，我們使用即時聚合酶連鎖反應系統 (real-time PCR)，量化量測施以外力時間長短造成的骨細胞基質基因調控變化，並分析其訊號傳遞機制。

　　換言之，本論文為細胞生物力學發展及整合許多研究技術，不僅有助于理解細胞力學訊息傳遞，而且可為組織工程和再生醫學提供更寬廣的應用。

　“Tissue Engineers” was been ranked as number 1 out of the ten hottest jobs in 21st century due to the brakethrough in organ failure treatment and the shortage from current options of tissue loss. With integrated interdiscipline of molecular biology, biochemistry, material science, clinical medicine, and biomedical engineering, the tissue engineering has been shown to evolve toward a powerful new paradigm of functional restoration or regeneration of lost or degenerated tissues and organs. While the tissue and cell remodeling is known to involve with the mechanical factors, the way how the cell senses the mechanical force and transduces the signal into gene regulation for remodeling become an important issue for regeneration medicine.

　The advance of the modern micro-electro-mechanical system (MEMS) and nanotechnologies has made the manufacturing scale to be able to be achieved to micrometer (µm, 10-6 meters) and even nanometer (nm, 10-9 meters) which is closely to the cellular or sub-cellular levels. Utilizing Bio-MEMS and nano-optical technologies, we established the manipulation and detection techniques for understanding how the cell senses the mechanical stimulation and transduces the intra- and intercellular signal to restore the tissue function.

　In cell manipulation, we utilized a custom optical tweezers and a self-developed cytodetachment system to quantitatively measure cell adhesion force at each different stage of cell’s life cycle, spreading and migration. We further used microlithography techniques to create different geometrical restriction of cell shape by micropatterning extracellular matrix (ECM) to investigate cell remodeling by application of an external force in different directions.

　For real-time measures of sub-cellular activities, the total internal reflection fluorescent microscopy (TIRFM) and fluorescent resonance energy transfer (FRET) systems were established for investigating the interaction between cell focal adhesion (FAK), phosphorylation of adhesion related molecule (Src), and cytoskeleton remodeling in response to external forces.

　The real-time polymerase chain reaction (PCR) technique was used to monitor the gene regulation during application of the external force, then the mechanism of mechanical sensing, mechanotransduction, and cell remodeling was investigated by protein activities analysis.

　In summary, this thesis described our development and integration of several novel technologies and skills for studies of cellular biomechanics to provide not only the understanding of mechanotransduction, but also broader implication in the regeneration medicine.

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